EP2358301B1 - Greffes de tissu régénératives et leurs procédés de fabrication - Google Patents
Greffes de tissu régénératives et leurs procédés de fabrication Download PDFInfo
- Publication number
- EP2358301B1 EP2358301B1 EP09829422.6A EP09829422A EP2358301B1 EP 2358301 B1 EP2358301 B1 EP 2358301B1 EP 09829422 A EP09829422 A EP 09829422A EP 2358301 B1 EP2358301 B1 EP 2358301B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- mpcs
- tissue
- nerve
- graft
- cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Not-in-force
Links
- 238000000034 method Methods 0.000 title description 42
- 230000001172 regenerating effect Effects 0.000 title description 11
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims description 199
- 210000005036 nerve Anatomy 0.000 claims description 85
- 210000001519 tissue Anatomy 0.000 claims description 71
- 239000000835 fiber Substances 0.000 claims description 70
- 208000027418 Wounds and injury Diseases 0.000 claims description 57
- 208000014674 injury Diseases 0.000 claims description 47
- 239000011159 matrix material Substances 0.000 claims description 47
- 230000006378 damage Effects 0.000 claims description 45
- 230000007547 defect Effects 0.000 claims description 33
- 230000008929 regeneration Effects 0.000 claims description 32
- 238000011069 regeneration method Methods 0.000 claims description 32
- 210000000130 stem cell Anatomy 0.000 claims description 32
- 239000002121 nanofiber Substances 0.000 claims description 29
- 210000000988 bone and bone Anatomy 0.000 claims description 21
- 239000012530 fluid Substances 0.000 claims description 18
- 230000001413 cellular effect Effects 0.000 claims description 17
- 230000035876 healing Effects 0.000 claims description 16
- 230000001225 therapeutic effect Effects 0.000 claims description 11
- 230000008439 repair process Effects 0.000 claims description 9
- 230000001737 promoting effect Effects 0.000 claims description 7
- 230000002792 vascular Effects 0.000 claims description 6
- 208000028389 Nerve injury Diseases 0.000 claims description 5
- 239000013543 active substance Substances 0.000 claims description 5
- 239000003102 growth factor Substances 0.000 claims description 5
- 230000008764 nerve damage Effects 0.000 claims description 5
- 239000003242 anti bacterial agent Substances 0.000 claims description 4
- 229940088710 antibiotic agent Drugs 0.000 claims description 4
- 230000000717 retained effect Effects 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 229940121363 anti-inflammatory agent Drugs 0.000 claims description 2
- 239000002260 anti-inflammatory agent Substances 0.000 claims description 2
- 230000003110 anti-inflammatory effect Effects 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000003443 antiviral agent Substances 0.000 claims 1
- 229940121357 antivirals Drugs 0.000 claims 1
- 239000006143 cell culture medium Substances 0.000 claims 1
- 239000012209 synthetic fiber Substances 0.000 claims 1
- 229920002994 synthetic fiber Polymers 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 94
- 210000003205 muscle Anatomy 0.000 description 45
- 230000012010 growth Effects 0.000 description 23
- 230000004069 differentiation Effects 0.000 description 22
- 230000014509 gene expression Effects 0.000 description 20
- 108090000623 proteins and genes Proteins 0.000 description 20
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 20
- 108010025020 Nerve Growth Factor Proteins 0.000 description 19
- 206010052428 Wound Diseases 0.000 description 19
- 210000001185 bone marrow Anatomy 0.000 description 19
- 239000000463 material Substances 0.000 description 19
- 239000002609 medium Substances 0.000 description 18
- 230000003376 axonal effect Effects 0.000 description 17
- 230000001228 trophic effect Effects 0.000 description 17
- 102000007072 Nerve Growth Factors Human genes 0.000 description 16
- 230000006698 induction Effects 0.000 description 16
- 239000003900 neurotrophic factor Substances 0.000 description 16
- 210000000578 peripheral nerve Anatomy 0.000 description 16
- 230000008569 process Effects 0.000 description 15
- 210000003050 axon Anatomy 0.000 description 14
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 12
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 239000000523 sample Substances 0.000 description 12
- 230000000508 neurotrophic effect Effects 0.000 description 11
- 230000002188 osteogenic effect Effects 0.000 description 11
- 229930182555 Penicillin Natural products 0.000 description 10
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 10
- APKFDSVGJQXUKY-INPOYWNPSA-N amphotericin B Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/C=C/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 APKFDSVGJQXUKY-INPOYWNPSA-N 0.000 description 10
- 229940049954 penicillin Drugs 0.000 description 10
- 229960005322 streptomycin Drugs 0.000 description 10
- 238000001804 debridement Methods 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 9
- 238000002955 isolation Methods 0.000 description 9
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 102100038169 Musculin Human genes 0.000 description 8
- 210000004271 bone marrow stromal cell Anatomy 0.000 description 8
- 238000001523 electrospinning Methods 0.000 description 8
- 210000002894 multi-fate stem cell Anatomy 0.000 description 8
- 238000001356 surgical procedure Methods 0.000 description 8
- 102100022464 5'-nucleotidase Human genes 0.000 description 7
- 239000012981 Hank's balanced salt solution Substances 0.000 description 7
- 101000678236 Homo sapiens 5'-nucleotidase Proteins 0.000 description 7
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 7
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- -1 CTNF Proteins 0.000 description 6
- 206010017076 Fracture Diseases 0.000 description 6
- 101000622304 Homo sapiens Vascular cell adhesion protein 1 Proteins 0.000 description 6
- 102100023543 Vascular cell adhesion protein 1 Human genes 0.000 description 6
- 238000004891 communication Methods 0.000 description 6
- 239000002131 composite material Substances 0.000 description 6
- 230000029087 digestion Effects 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 230000005012 migration Effects 0.000 description 6
- 238000013508 migration Methods 0.000 description 6
- 230000011164 ossification Effects 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 210000002435 tendon Anatomy 0.000 description 6
- 238000002560 therapeutic procedure Methods 0.000 description 6
- 208000010392 Bone Fractures Diseases 0.000 description 5
- 102100037241 Endoglin Human genes 0.000 description 5
- 101000881679 Homo sapiens Endoglin Proteins 0.000 description 5
- 241000906034 Orthops Species 0.000 description 5
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 239000003636 conditioned culture medium Substances 0.000 description 5
- 239000012091 fetal bovine serum Substances 0.000 description 5
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 210000002241 neurite Anatomy 0.000 description 5
- 229920000642 polymer Polymers 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 229930002330 retinoic acid Natural products 0.000 description 5
- 210000004116 schwann cell Anatomy 0.000 description 5
- 230000017423 tissue regeneration Effects 0.000 description 5
- 230000008733 trauma Effects 0.000 description 5
- 229960001727 tretinoin Drugs 0.000 description 5
- 239000012583 B-27 Supplement Substances 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 102100022640 Collagen alpha-1(XV) chain Human genes 0.000 description 4
- 102100040895 Growth/differentiation factor 10 Human genes 0.000 description 4
- 208000034970 Heterotopic Ossification Diseases 0.000 description 4
- 101000899935 Homo sapiens Collagen alpha-1(XV) chain Proteins 0.000 description 4
- 101000893563 Homo sapiens Growth/differentiation factor 10 Proteins 0.000 description 4
- 101001076292 Homo sapiens Insulin-like growth factor II Proteins 0.000 description 4
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 description 4
- 102100025947 Insulin-like growth factor II Human genes 0.000 description 4
- 102000000536 PPAR gamma Human genes 0.000 description 4
- 229920000954 Polyglycolide Polymers 0.000 description 4
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 238000002659 cell therapy Methods 0.000 description 4
- 210000002889 endothelial cell Anatomy 0.000 description 4
- 230000007941 heterotopic ossification Effects 0.000 description 4
- 230000001506 immunosuppresive effect Effects 0.000 description 4
- 239000007943 implant Substances 0.000 description 4
- 230000001965 increasing effect Effects 0.000 description 4
- 230000001338 necrotic effect Effects 0.000 description 4
- 210000000963 osteoblast Anatomy 0.000 description 4
- 230000009818 osteogenic differentiation Effects 0.000 description 4
- 239000004633 polyglycolic acid Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 230000035899 viability Effects 0.000 description 4
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 102000009123 Fibrin Human genes 0.000 description 3
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 3
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 3
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 3
- 102000015336 Nerve Growth Factor Human genes 0.000 description 3
- 102100029268 Neurotrophin-3 Human genes 0.000 description 3
- 102000004067 Osteocalcin Human genes 0.000 description 3
- 108090000573 Osteocalcin Proteins 0.000 description 3
- 108091008767 PPARγ2 Proteins 0.000 description 3
- 210000001789 adipocyte Anatomy 0.000 description 3
- 210000004504 adult stem cell Anatomy 0.000 description 3
- 230000028600 axonogenesis Effects 0.000 description 3
- 229920000249 biocompatible polymer Polymers 0.000 description 3
- 210000000845 cartilage Anatomy 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 230000001143 conditioned effect Effects 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 210000003195 fascia Anatomy 0.000 description 3
- 229950003499 fibrin Drugs 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000003306 harvesting Methods 0.000 description 3
- 238000002513 implantation Methods 0.000 description 3
- 210000003041 ligament Anatomy 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 238000010172 mouse model Methods 0.000 description 3
- 230000001537 neural effect Effects 0.000 description 3
- 238000010899 nucleation Methods 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 238000007747 plating Methods 0.000 description 3
- 238000002203 pretreatment Methods 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 239000002002 slurry Substances 0.000 description 3
- 230000008093 supporting effect Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 210000005166 vasculature Anatomy 0.000 description 3
- 239000011800 void material Substances 0.000 description 3
- 230000029663 wound healing Effects 0.000 description 3
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 3
- ZAJGKKXBKLVSKW-UHFFFAOYSA-N 3,6-dimethyl-1,4-dioxane-2,5-dione;oxepan-2-one Chemical compound O=C1CCCCCO1.CC1OC(=O)C(C)OC1=O ZAJGKKXBKLVSKW-UHFFFAOYSA-N 0.000 description 2
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- 101710151806 72 kDa type IV collagenase Proteins 0.000 description 2
- 102100026802 72 kDa type IV collagenase Human genes 0.000 description 2
- 102100032912 CD44 antigen Human genes 0.000 description 2
- 108010005939 Ciliary Neurotrophic Factor Proteins 0.000 description 2
- 102100031614 Ciliary neurotrophic factor Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- 102000029816 Collagenase Human genes 0.000 description 2
- 108060005980 Collagenase Proteins 0.000 description 2
- 241000283074 Equus asinus Species 0.000 description 2
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 102000034615 Glial cell line-derived neurotrophic factor Human genes 0.000 description 2
- 108091010837 Glial cell line-derived neurotrophic factor Proteins 0.000 description 2
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 2
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 2
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 2
- 101000994369 Homo sapiens Integrin alpha-5 Proteins 0.000 description 2
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 2
- 206010062016 Immunosuppression Diseases 0.000 description 2
- 102100032817 Integrin alpha-5 Human genes 0.000 description 2
- 102000002274 Matrix Metalloproteinases Human genes 0.000 description 2
- 108010000684 Matrix Metalloproteinases Proteins 0.000 description 2
- 108010015302 Matrix metalloproteinase-9 Proteins 0.000 description 2
- 102100030412 Matrix metalloproteinase-9 Human genes 0.000 description 2
- 102000008730 Nestin Human genes 0.000 description 2
- 108010088225 Nestin Proteins 0.000 description 2
- 208000010886 Peripheral nerve injury Diseases 0.000 description 2
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 2
- 239000004793 Polystyrene Substances 0.000 description 2
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- 238000000692 Student's t-test Methods 0.000 description 2
- 208000002847 Surgical Wound Diseases 0.000 description 2
- 230000006052 T cell proliferation Effects 0.000 description 2
- 102000056172 Transforming growth factor beta-3 Human genes 0.000 description 2
- 108090000097 Transforming growth factor beta-3 Proteins 0.000 description 2
- 230000002293 adipogenic effect Effects 0.000 description 2
- 210000000577 adipose tissue Anatomy 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000001733 anti-osteogenic effect Effects 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 229920002988 biodegradable polymer Polymers 0.000 description 2
- 239000004621 biodegradable polymer Substances 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 230000037186 bone physiology Effects 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 230000035605 chemotaxis Effects 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 230000002648 chondrogenic effect Effects 0.000 description 2
- 210000003040 circulating cell Anatomy 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 229960002424 collagenase Drugs 0.000 description 2
- 230000001332 colony forming effect Effects 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 230000009274 differential gene expression Effects 0.000 description 2
- 230000003467 diminishing effect Effects 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 210000003414 extremity Anatomy 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 230000002518 glial effect Effects 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 238000013394 immunophenotyping Methods 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000028709 inflammatory response Effects 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002062 molecular scaffold Substances 0.000 description 2
- 230000009756 muscle regeneration Effects 0.000 description 2
- 230000007336 muscle tissue development Effects 0.000 description 2
- 230000004070 myogenic differentiation Effects 0.000 description 2
- 230000001114 myogenic effect Effects 0.000 description 2
- 210000005055 nestin Anatomy 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000001582 osteoblastic effect Effects 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 229920005594 polymer fiber Polymers 0.000 description 2
- 229920002223 polystyrene Polymers 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002062 proliferating effect Effects 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 210000004872 soft tissue Anatomy 0.000 description 2
- 210000002536 stromal cell Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 230000008736 traumatic injury Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 230000037314 wound repair Effects 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- PJOHVEQSYPOERL-SHEAVXILSA-N (e)-n-[(4r,4as,7ar,12br)-3-(cyclopropylmethyl)-9-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1h-4,12-methanobenzofuro[3,2-e]isoquinoline-4a-yl]-3-(4-methylphenyl)prop-2-enamide Chemical compound C1=CC(C)=CC=C1\C=C\C(=O)N[C@]1(CCC(=O)[C@@H]2O3)[C@H]4CC5=CC=C(O)C3=C5[C@]12CCN4CC1CC1 PJOHVEQSYPOERL-SHEAVXILSA-N 0.000 description 1
- 102100024506 Bone morphogenetic protein 2 Human genes 0.000 description 1
- 102100024505 Bone morphogenetic protein 4 Human genes 0.000 description 1
- 102100022525 Bone morphogenetic protein 6 Human genes 0.000 description 1
- 102100023126 Cell surface glycoprotein MUC18 Human genes 0.000 description 1
- 102000007644 Colony-Stimulating Factors Human genes 0.000 description 1
- 108010071942 Colony-Stimulating Factors Proteins 0.000 description 1
- 102000015775 Core Binding Factor Alpha 1 Subunit Human genes 0.000 description 1
- 108010024682 Core Binding Factor Alpha 1 Subunit Proteins 0.000 description 1
- 102100036912 Desmin Human genes 0.000 description 1
- 108010044052 Desmin Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical group CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 102100028412 Fibroblast growth factor 10 Human genes 0.000 description 1
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 102100035368 Growth/differentiation factor 6 Human genes 0.000 description 1
- 101000762366 Homo sapiens Bone morphogenetic protein 2 Proteins 0.000 description 1
- 101000762379 Homo sapiens Bone morphogenetic protein 4 Proteins 0.000 description 1
- 101000899390 Homo sapiens Bone morphogenetic protein 6 Proteins 0.000 description 1
- 101000623903 Homo sapiens Cell surface glycoprotein MUC18 Proteins 0.000 description 1
- 101000917237 Homo sapiens Fibroblast growth factor 10 Proteins 0.000 description 1
- 101001023964 Homo sapiens Growth/differentiation factor 6 Proteins 0.000 description 1
- 101001023043 Homo sapiens Myoblast determination protein 1 Proteins 0.000 description 1
- 101001086210 Homo sapiens Osteocalcin Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102000003814 Interleukin-10 Human genes 0.000 description 1
- 108090000174 Interleukin-10 Proteins 0.000 description 1
- 108010085895 Laminin Proteins 0.000 description 1
- 101150110301 MSC gene Proteins 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102100035077 Myoblast determination protein 1 Human genes 0.000 description 1
- 208000002565 Open Fractures Diseases 0.000 description 1
- 102100031475 Osteocalcin Human genes 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108010016731 PPAR gamma Proteins 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 238000010240 RT-PCR analysis Methods 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 208000026062 Tissue disease Diseases 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000009815 adipogenic differentiation Effects 0.000 description 1
- 238000002266 amputation Methods 0.000 description 1
- 229940035676 analgesics Drugs 0.000 description 1
- 239000000730 antalgic agent Substances 0.000 description 1
- 230000003510 anti-fibrotic effect Effects 0.000 description 1
- 230000002921 anti-spasmodic effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 229940124575 antispasmodic agent Drugs 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 230000003416 augmentation Effects 0.000 description 1
- 230000004009 axon guidance Effects 0.000 description 1
- 230000010442 axonal sprouting Effects 0.000 description 1
- 239000012867 bioactive agent Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000002798 bone marrow cell Anatomy 0.000 description 1
- 230000010478 bone regeneration Effects 0.000 description 1
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 description 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 1
- 230000003848 cartilage regeneration Effects 0.000 description 1
- 239000002458 cell surface marker Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000011198 co-culture assay Methods 0.000 description 1
- 229940047120 colony stimulating factors Drugs 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000003271 compound fluorescence assay Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011257 definitive treatment Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 210000005045 desmin Anatomy 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 1
- 239000010432 diamond Substances 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000003292 diminished effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000000635 electron micrograph Methods 0.000 description 1
- 230000010595 endothelial cell migration Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 230000001973 epigenetic effect Effects 0.000 description 1
- 210000000267 erythroid cell Anatomy 0.000 description 1
- 230000000925 erythroid effect Effects 0.000 description 1
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 1
- 229960005542 ethidium bromide Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 238000007804 gelatin zymography Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 229940125721 immunosuppressive agent Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 238000012977 invasive surgical procedure Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000007799 mixed lymphocyte reaction assay Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000001565 modulated differential scanning calorimetry Methods 0.000 description 1
- 230000010070 molecular adhesion Effects 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 230000000921 morphogenic effect Effects 0.000 description 1
- 229940035363 muscle relaxants Drugs 0.000 description 1
- 210000001665 muscle stem cell Anatomy 0.000 description 1
- 230000023105 myelination Effects 0.000 description 1
- 210000004985 myeloid-derived suppressor cell Anatomy 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 239000003158 myorelaxant agent Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229940053128 nerve growth factor Drugs 0.000 description 1
- 210000000944 nerve tissue Anatomy 0.000 description 1
- 210000004498 neuroglial cell Anatomy 0.000 description 1
- 229960002378 oftasceine Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000004072 osteoblast differentiation Effects 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 210000003668 pericyte Anatomy 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- RLZZZVKAURTHCP-UHFFFAOYSA-N phenanthrene-3,4-diol Chemical compound C1=CC=C2C3=C(O)C(O)=CC=C3C=CC2=C1 RLZZZVKAURTHCP-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920001610 polycaprolactone Polymers 0.000 description 1
- 239000004626 polylactic acid Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000001023 pro-angiogenic effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000002278 reconstructive surgery Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000007832 reinnervation Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 230000036573 scar formation Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000000405 serological effect Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 210000002027 skeletal muscle Anatomy 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 210000005222 synovial tissue Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000008467 tissue growth Effects 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 238000011541 total hip replacement Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000000472 traumatic effect Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000003954 umbilical cord Anatomy 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/28—Materials for coating prostheses
- A61L27/34—Macromolecular materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/04—Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts
- A61F2/06—Blood vessels
- A61F2/062—Apparatus for the production of blood vessels made from natural tissue or with layers of living cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2210/00—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2210/0004—Particular material properties of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof bioabsorbable
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2250/00—Special features of prostheses classified in groups A61F2/00 - A61F2/26 or A61F2/82 or A61F9/00 or A61F11/00 or subgroups thereof
- A61F2250/0058—Additional features; Implant or prostheses properties not otherwise provided for
- A61F2250/0067—Means for introducing or releasing pharmaceutical products into the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K2035/124—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells the cells being hematopoietic, bone marrow derived or blood cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/12—Nanosized materials, e.g. nanofibres, nanoparticles, nanowires, nanotubes; Nanostructured surfaces
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/40—Nucleotides, nucleosides, bases
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/44—Thiols, e.g. mercaptoethanol
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/01—Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/13—Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/235—Leukemia inhibitory factor [LIF]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/33—Insulin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/38—Hormones with nuclear receptors
- C12N2501/385—Hormones with nuclear receptors of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/38—Hormones with nuclear receptors
- C12N2501/39—Steroid hormones
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
- C12N2533/40—Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
Definitions
- Debridement of contaminated and devitalized tissue is the first step in the surgical treatment of open extremity injuries. This event is often part of a process comprising serial debridements over the span of several days to fully assess the viability of the remaining tissue.
- Penetrating trauma results in substantial bone and soft tissue loss due to the primary injury and the debridement process.
- a projectile or blast wave penetrates the skin, it transfers kinetic energy to the surrounding structures, which include bone, muscle, tendon, cartilage, and fat.
- Jussila J Measurement of kinetic energy dissipation with gelatine fissure formation with special reference to gelatine validation. Forensic Sci Int. 2005, 150: 53-62 . This energy is absorbed in the form of heat, mechanical stress, and chemical stress, and it initiates a number of events, including cell necrosis, apoptosis, and inflammation.
- tissue-grafting or tissue substitutes that are often derived from allograft or synthetic sources.
- revision and reconstructive surgery is frequently required to restore the function of the injured extremity. In many instances, this may require bone and soft-tissue augmentation, lysis of adhesions (about joints and along tendons), and/or ligament reconstruction.
- revision surgery stems from a need to repair or replace absent, damaged, or deranged tissues such as articular cartilage, tendon, and/or bone with use of autograft, allograft, bioengineered tissue replacement, or prosthetic materials and devices.
- these options for tissue repair or replacement are limited by the inability of the implant to fully integrate and subsequently remodel.
- the inferior structural, biomechanical, and biochemical properties of the implant as compared with normal human tissue prevent full restoration of the structure-function relationship.
- tissue- engineering construct designs An essential component of all tissue- engineering construct designs is a readily available, viable, and plastic cell source.
- Many sources of multipotent progenitor cells e.g., bone marrow, trabecular bone, adipose tissue, umbilical cord blood, and synovial tissue
- Caterson EJ, et al Human marrow-derived mesenchymal progenitor cells: isolation, culture expansion, and analysis of differentiation. Mol Biotechnol., 2002, 20:245-56 ; Noth U, et al, Multilineage mesenchymal differentiation potential of human trabecular bone-derived cells.
- Peripheral nerve injury frequently accompanies musculoskeletal trauma, which lengthens the recovery time and leads to significant dysfunction.
- Current treatment of peripheral nerve injuries includes primary repair, nerve autograft, or use of synthetic nerve tubes. The success of nerve repair depends primarily on the speed of axonal growth and myelination to bridge the damaged region and decrease the time to end organ re-innervation.
- Electrospun tubes with no additional biological coating or drug loading treatment, may be used as scaffolds for functional nervous regeneration.
- Silvia Panseri et al. "Electrospun micro- and nanofiber tubes for functional nervous regeneration in sciatic nerve transections.”, BMC BIOTECHNOLOGY, vol. 8, 11 April 2008 .
- Conventional nerve tubes contain a single lumen to guide the regenerating nerve from proximal to distal stump. Although increasing the likelihood that some axons in the nerve will reconnect with the distal end, many are unable to reconnect, and gaps larger than critical size defect are likely never to regenerate.
- MPCs Mesenchymal progenitor cells
- MPCs Mesenchymal progenitor cells
- MPCs mesenchymal progenitor cells
- MPCs offer advantages over MSCs of being more plentiful (particularly in wound tissue), more easily obtained, and capable of producing substantial quantity of various trophic factors, including neurotrophic factors, vasculotrophic factors, and osteotrophic factors.
- MPCs have the added advantage of diminishing inflammation and scar formation.
- MPCs in the treatment of various tissue injury or disease states.
- the MPCs can be used as an in situ or in vivo source of various biologically significant and therapeutically effective factors, e.g., growth factors and differentiation inducing factors that promote regeneration and/or healing.
- the MPCs are not consumed, nor are they completely differentiated in the process.
- Jackson, W.M, et al. Putative Heterotopic Ossification Progenitor Cells Derived from Traumatized Muscle, J. Ortho. Research, 1, (2009) (published online at www.interscience.wiley.com, DOI 10.1002/jor.20924 ).
- MPCs substantially remain as MPCs and continue to secrete trophic factors and promote differentiation of other cell types to regenerate the damaged tissue for a considerable time.
- MPCs In vitro studies show that MPCs continue to secrete trophic factors when the MPCs were at passage 2-3, which corresponds roughly to 12-15 population doublings, or approximately 3-4 weeks after harvest. MPCs likely continue to express trophic factors as long as they remain in an undifferentiatied state.
- the grafts of the present invention exploit unique properties of MPCs, i.e., in a wound setting they remain in an undifferentiated state for a prolonged period, and during that time secrete trophic factors that aid in the regeneration of nerve, bone, vasculature and the like.
- the present invention provides a graft for promoting healing of an injury or defect comprising a scaffold and a therapeutic component, the scaffold comprising a porous matrix of non-aligned fibers forming interstices, wherein the therapeutic component comprises a cellular component comprising mesenchymal progenitor cells and, optionally, a non-cellular component, wherein the mesenchymal progenitor cells are retained in the interstices formed by the non-aligned fibers of the porous matrix of the graft in a manner that places the mesenchymal progenitor cells in proximity to, and in fluid contact with, but outside the injury or defect and avoids contact of the mesenchymal progenitor cells to the site of the injury or defect, wherein the graft has the form of a conduit comprising an inner sheath of aligned nanofibers that is surrounded by the non-aligned fibers forming an outer sheath.
- the matrix serves as a reservoir or repository for
- the therapeutic component may include other therapeutic elements such as small molecule active agents commonly used to aid healing or fight infection.
- the matrix retains the therapeutic component such as the MPCs, but permits fluid flow within and through the matrix.
- the MPC-seeded matrix may be placed proximate to the injury or defect, and in fluid contact with the injury or defect.
- the term matrix is used to refer to a porous material within which MPCs, and perhaps other cells or therapeutic materials, may be infused and retained.
- the matrix material may be natural or synthetic, and may be additionally treated with substances to enhance retention of cells, e.g., treatment of polymeric material with hyaluronic acid. While it is desirable to retain MPCs and the like within the matrix, it is to be understood that some migration will occur, both to the interior and exterior of the various grafts.
- the porosity and retention capacity of the matrix can be varied according to known methods depending on the intended application, the type of graft employed, the type of tissue under treatment, and the severity of the injury or defect.
- a second scaffold component may be added to the graft.
- One such element provides structural support for tissue regeneration and/or isolation and protection of the injury or defect.
- the second component can be a structural element serving as a guide or framework on which the regenerated tissue can form, or it can be a structural element that creates and/or retains a void in or around the defect, or otherwise supports and/or retains the damaged tissue as it is to be reformed so as to permit the tissue to regenerate and return to its original dimension and shape.
- tissue includes nerve, bone, and vascular tissue.
- These grafts may also comprise an element exterior to the matrix to isolate and/or concentrate the MPCs and the various factors produced, and to avoid loss or migration of other regenerative elements such as other cell types.
- the exterior element can be fabricated to match the overall structure of the scaffold, i.e., a conduit.
- the material used in all of the foregoing elements may be the same or different, and may be fabricated to afford differing levels of porosity.
- the exterior element can be fabricated to be substantially or completely non-porous.
- the exterior element which may function as a dam or fluid retention device to protect and isolate therapeutic components, can also be formed of biocompatible nanofibers, and created to varying levels of porosity.
- the dam can be made substantially or entirely fluid impermeable; or it can be made to be permeable but less so than the matrix.
- MPCs express neurotrophic factors (e.g., BDNF, CTNF, NT-3), which encourage axonal growth and nerve regeneration. Since nerve damage frequently occurs in orthopaedic injury, the traumatized muscle-derived progenitor cells are a readily available autologous cell source that can be utilized to effect or improve nerve repair. MPCs may be used quite effectively in a nerve graft.
- neurotrophic factors e.g., BDNF, CTNF, NT-3
- a device for regenerating injured or damaged nerve, and for enhancing the rate of axonal growth incorporates a composite of nanofiber structures, and wherein the fibers are used to construct two zones or chambers within the nerve graft conduit.
- a core of aligned fibers In the interior of the graft is a core of aligned fibers along the axis of symmetry through the longitudinal axis of symmetry of the graft.
- An exterior sheath surrounding the core is fabricated of randomly non-aligned fibers to serve as a matrix for seeding and supporting the MPCs, which promotes the activity of the MPCs and other endogenous neuronal support cells (e.g., Schwann Cells can migrate into the graft).
- endogenous neuronal support cells e.g., Schwann Cells can migrate into the graft.
- a novel peripheral nerve graft to include a core of aligned nanometer-scale fibers, and an outer sheath to support and retain the MPCs proximate to the wound or injury following implantation of the device.
- the aligned fibers guide the regeneration of individual neurons of the nerve.
- MPCs are loaded into a nerve graft in the form of a tissue engineered peripheral nerve conduit.
- One such conduit includes a porous matrix in the form of an outer sheath. MPCs are seeded within the matrix. Trophic factors produced by MPCs are secreted within close proximity of the injured nerve tissue thereby promoting healing and differentiation of other cell types to regenerate the damaged tissue.
- the outer sheath surrounds an interior area of the conduit.
- the interior area may be a void within which the nerve regeneration is afforded space to reconnect the proximal and distal ends of the nerve stub, or it may comprise a core of aligned nanometer-scale fibers that guide the regeneration of individual neurons of the nerve.
- the aligned fibers may be generally linear along an axis of symmetry running longitudinally through the conduit. Trophic factors produced by MPCs diffuse to the region of the aligned fibers, and promote regeneration of the nerve along those fibers.
- a nerve graft in one embodiment, there is a nerve graft, and methods of making same, wherein the graft comprises a nerve tube having a central region or sheath filled with aligned fibers (also referred to herein as "nanofibers").
- aligned fibers also referred to herein as "nanofibers”
- nanofibers refers to fibers of about 0.05-0.5 ⁇ m in diameter; or about 0.1-0.3 ⁇ m in diameter; or 0.2 ⁇ m in diameter.
- the nerve tube comprises a composite of electrospun fibers, both linear (e.g., at the inner sheath) and nonlinear (e.g., at the outer sheath).
- the nerve tube device promotes axonal growth along the linear, aligned fibers aided by neurotrophic factors secreted by support cells seeded in the support matrix.
- the nerve tube is fabricated by a novel two step electrospinning process. The novel electrospinning process can produce electrospun fibers on a non-conductive mandrel.
- Axonal nerve tubes have previously been used to guide the regeneration of damaged nerve.
- Commercially available nerve tubes consist of collagen, or a similar biological or biocompatible polymer, in the shape of a hollow tube that is telescoped over the ends of the damaged nerve stumps.
- the limitation of these devices is related to the speed of the axonal growth through the hollow interior of the tube. If the axons do not reconnect with the distal stump within several weeks, the nerve cannot be regenerated.
- a device that increases rate of axonal growth will bridge longer nerve gaps during axonal growth.
- the devices and methods disclosed herein facilitate functional regeneration for a wide range of nerve injuries, both in terms of length of damaged nerve and in speed of regeneration.
- MPCs express neurotrophic factors (e.g., BDNF, CTNF, NT-3).
- the neurotrophic factors stimulate and promote axonal growth and nerve regeneration by increasing the rate of axonal outgrowth. Since nerve damage frequently occurs in orthopaedic injury, the traumatized muscle-derived progenitor cells are a readily available autologous cell source that improve nerve repair. This can be achieved by loading MPCs into a tissue engineered peripheral nerve conduit (nerve tube) as described elsewhere herein.
- the present invention affords means and devices for using a new cell type (MPC) found in traumatized muscle tissue.
- MPCs possess potent regenerative properties. They can be harvested in high numbers directly from traumatized muscle tissue, and can be employed immediately in various therapies for effecting nerve regeneration. MPCs afford an autologous resource for therapeutic material useful in various new and known therapies. By affording an autologous resource, the risk of allogenic response in a patient is dramatically reduced, if not eliminated.
- Debrided muscle contains multipotent cells useful in cell-based tissue-engineering therapies and studies. Following a modified stem cell isolation protocol on tissues obtained at surgery, we obtained viable cells expressing markers characteristic of mesenchymal stem cells. See, e.g., Chamberlain G, et al, Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing, Stem Cells, 2007, 25:2739-49 ; Baksh D, et al, Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow, Stem Cells, 2007, 25:1384-92 . We defined these cells as mesenchymal progenitor cells, or MPCs.
- MPCs mesenchymal progenitor cells were culture-expanded, and they exhibited multipotentiality (adipogenic, osteogenic, and chondrogenic) on appropriate induction. These MPCs can be used in the initial reparative process or in future reconstructive operations in combination with appropriate tissue-engineering biomaterial scaffolds.
- the differentiation assays were corroborated by the expression of corresponding adipogenic, osteogenic, and chondrogenic lineage-specific genes.
- muscle-derived MPCs have the potential to differentiate into osteoblasts, adipocytes, and chondrocytes.
- MPCs derived from traumatized muscle require substantial characterization in terms of their origin within the body and their relationship to better-characterized stem cell types.
- the MPCs may originally reside in the nontraumatized muscle tissue in a quiescent state (i.e., as pericytes) ( Bianco P, et al, Mesenchymal stem cells: revisiting history, concepts, and assays, Cell Stem Cell., 2008, 2:313-9 ), or they may have migrated from the bone marrow to the site of injury in response to wound-healing signals.
- Kumagai K, et al Circulating cells with osteogenic potential are physiologically mobilized into the fracture healing site in the parabiotic mice model.
- PPAR ⁇ 2 which is an indicator of adipogenic differentiation in bone marrow-derived mesenchymal stem cells, is also upregulated by osteogenic induction of traumatized muscle-derived mesenchymal progenitor cells.
- the effect of PPAR ⁇ 2 on the MPCs does not appear to be anti-osteogenic as there is strong evidence from the histological and gene-expression findings that cells underwent osteogenic induction.
- the regulatory pathways governing PPAR ⁇ 2 activity can modulate its anti-osteogenic function ( Lecka-Czemik B, et al, Divergent effects of selective peroxisome proliferator-activated receptor gamma 2 ligands on adipocyte versus osteoblast differentiation, Endocrinology, 2002, 143:2376-84 ) and may represent a tissue-specific feature of regenerative cells that are present in muscle tissue.
- Surgical debridement of open wounds is a medical and surgical necessity. Although removal of tissue from wounds that are characterized by substantial tissue loss is counterintuitive, it is essential for definitive treatment, wound closure, and proper healing. Granick M, et al, Toward a common language: surgical wound bed preparation and debridement. Wound Repair Regen, 2006, 14 Suppl 1:S1-10 ; Gregory P, et al, The management of severe fractures of the lower extremities. Clin Orthop Relat Res. 1995;318:95-105 ; Jacob E, et al, A retrospective analysis of open fractures sustained by U.S. military personnel during Operation Just Cause. Mil Med., 1992, 157:552-6 .
- Qu-Petersen and colleagues described the presence of a population of progenitor cells obtained from skeletal muscle in a mouse model that exhibited characteristics similar to, but distinct from, mesenchymal stem cells.
- Qu-Petersen Z, et al Identification of a novel population of muscle stem cells in mice: potential for muscle regeneration. J Cell Biol. 2002, 157:851-64 . They isolated a cell population by a preplating technique, which selects for the least-adherent cell population after a series of six serial platings. With this method, we were unable to obtain multiprogenitor cells from human tissue samples, and the cells that were obtained differed significantly from the multiprogenitor cells described here.
- MPCs express significantly greater levels of TGFB3; MPCs continue to proliferate while being induced to differentiate into osteoblasts, and express lower levels of osteocalcin, an osteoblastic gene that is expressed during later stages of osteogenic differentiation. Additionally, there are differences in the osteogenic gene expression profile between the MPCs and MSCs, which may reflect the tissue of origin for both cell types.
- MPCs express higher levels of COL15A1, a gene associated with muscle tissue development, and GDF10, shown to be a negative regulator of osteogenesis, whereas the bone-marrow derived MSCs express higher levels of genes associated with bone physiology and maintenance: VEGFA 17, VCAM1 18 and IGF2 19. These differences may also reflect the fact that traumatized muscle-derived MPCs are harvested from an active wound bed, where they likely participate in the process of muscle tissue repair. During osteogenic differentiation, COL15A1 and GDF10 are substantially down-regulated, while VEGFA, VCAM1 and IGF2 are similarly up-regulated, suggesting that the MPCs can assume the role of osteoprogenitors under the appropriate biological environment, in a manner similar to other populations of MSCs.
- the traumatized muscle-derived MPCs are of particular benefit in blast trauma-induced injuries given the high prevalence of heterotopic ossification associated with such injuries.
- E.g., Potter BK, et al. Heterotopic ossification following traumatic and combat-related amputations. Prevalence, risk factors, and preliminary results of excision, J Bone Joint Surg Am. 2007, 89:476-86 .
- the osteogenic potential of these cells suggests their possible role in pathological processes that result in ectopic bone formation.
- Granick M, et al Toward a common language: surgical wound bed preparation and debridement, Wound Repair Regen. 2006, 14 Suppl 1:S1-10 .
- a method for isolating MPCs comprises: structural degradation or destruction (e.g., mincing or chopping) of a muscle sample from a wound (e.g., a debrided wound); suspension of the sample in a digestion medium; culturing and/or incubating the sample; and isolating MPCs.
- structural degradation or destruction e.g., mincing or chopping
- a method for isolating MPCs comprises: removal of unhealthy tissue (e.g., fat, fascia, damaged connective tissue, and necrotic tissue) from a healthy margin of muscle in a wound; isolation of a sample of the muscle; washing the muscle sample; structural degradation or destruction (e.g., mincing or chopping) of the muscle sample to create a tissue suspension; washing the tissue suspension of the sample in a digestion medium; and culturing and/or incubating the sample; and isolating MPCs.
- the tissue may be repeatedly washed in a salt solution (e.g., Hanks' Balanced Salt Solution (Gibco, Carlsbad, California)).
- the tissue sample may be chopped or minced in digestion medium (e.g., Dulbecco's Modified Eagle Medium (Gibco)), to which antibiotic may be added (e.g., penicillin/streptomycin/Fungizone (Gibco)). Mincing or chopping of the sample is carried out until an appropriate particle size is achieved, e.g., about 5 mm 3 or less, or about 1 mm 3 , or such that the product can pass through a pipette).
- digestion medium e.g., Dulbecco's Modified Eagle Medium (Gibco)
- antibiotic e.g., penicillin/streptomycin/Fungizone (Gibco)
- Mincing or chopping of the sample is carried out until an appropriate particle size is achieved, e.g., about 5 mm 3 or less, or about 1 mm 3 , or such that the product can pass through a pipette).
- the minced tissue can then be transferred to digestion medium, e.g., containing Dulbecco's Modified Eagle Medium, 3X penicillin/streptomycin/Fungizone, and 0.5 mg/mL collagenase type 2 (Worthington Biochemical, Lakewood, New Jersey)).
- digestion medium e.g., containing Dulbecco's Modified Eagle Medium, 3X penicillin/streptomycin/Fungizone, and 0.5 mg/mL collagenase type 2 (Worthington Biochemical, Lakewood, New Jersey)
- the tissue slurry can then be cultured and/or incubated. In one embodiment, the tissue slurry is agitated at about 37°C for two hours.
- the resulting digest may then be filtered (e.g., through a 40- ⁇ m cell strainer (Falcon)).
- the resulting digest may also be subjected to centrifugation.
- a pellet resulting from a centrifugation step may be resuspended (e.g., in growth medium such as Dulbecco's Modified Eagle Medium with 10% fetal bovine serum; Gibco) and 5X penicillin/streptomycin/Fungizone).
- growth medium such as Dulbecco's Modified Eagle Medium with 10% fetal bovine serum; Gibco
- 5X penicillin/streptomycin/Fungizone e.g., in growth medium such as Dulbecco's Modified Eagle Medium with 10% fetal bovine serum; Gibco
- the resulting digest and/or the resuspended pellet may be plated onto tissue culture and incubated.
- incubation is carried out at elevated temperature (e.g., >25°C; or >30°C; or about 37°C); and may be carried out in a CO 2 -humidified environment (e.g., 1-10% CO 2 ; or about 5% CO 2 ).
- the incubation may be performed in a cell incubator (e.g., for at least about one-half hour; or for about two hours).
- the culture may be washed with a biologically compatible medium (.e.g., with Hanks' Balanced Salt Solution).
- a fresh growth medium may be added with additional antibiotic (e.g., 3X penicillin, streptomycin, & Fungizone).
- additional antibiotic e.g., 3X penicillin, streptomycin, & Fungizone.
- the concentration of penicillin, streptomycin, & Fungizone is lowered to about 1X.
- the cell cultures may be routinely passaged at 80% to 90% confluence and split (e.g., 1:4).
- One such method deploys two chambers or tubes.
- the chambers may be configured in shape and size as a conventional centrifuge tube. See Figure 3 .
- a first tube is outfitted with a tissue grinding device, optionally near an opening of a tube.
- the grinder chops or minces MPC-containing tissue (e.g., to about 5 mm 3 or less, or about 1 mm 3 or less).
- Interior to the grinder within the chamber is a reservoir of a medium containing enzymes, growth factors, antibiotics, and other suitable cell-sustaining agents.
- those agents are suspended in a sterile saline solution, and the tissue fragments are enzymatically digested.
- the digestion may occur over an incubation period, optionally at elevated temperature (e.g., >25°C; or ⁇ 37°C).
- the digested tissue is processed through another chamber, device or tube.
- the second chamber is outfitted with a series of shear channels, which cause the digested tissue fragments to disperse.
- the resulting tissue fragments then are processed through a cell strainer, which isolates the MPCs.
- the isolated MPCs are then suitable for implantation and use in the various methods and devices disclosed elsewhere herein, and in other methods and therapies as would occur to one of ordinary skill in the art. Such methods and therapies can be readily adapted to be performed at the point-of-care.
- MPCs Progenitor cells harvested from traumatized muscle have several characteristics of MSC: 1) similar morphology, proliferation rate, cell surface markers and gene expression profile; 2) differentiation into osteoblasts, adipocytes and chondrocytes; and 3) immunosuppressive, pro-angiogenic, anti-fibrotic properties. Additionally, the differentiation potential of MPC populations is uniform between patients; and traumatized muscle-derived MPCs could be harvested clinically for use in regenerative medicine applications, e.g., cellular therapy and tissue engineering.
- MPCs produce various bioactive agents that promote healing and regeneration, such as growth factors or differentiation factors, and/or promote the migration of other healing influences to the site of the injury or defect such as other cell types (e.g., Schwann cells), or that promote the actual differentiation of other cell types into lines needed to regenerate or restore injured or damaged tissue.
- the grafts are configured to place MPCs in proximity to the injury or defect, and in fluid contact with the injury or defect such that the agents produced and/or secreted by MPCs produce the intended effect at or within the injury.
- the grafts differ from other grafts incorporating other multiprogenitor cells such as stem cells in that the instant grafts avoid or diminish access or contact of the MPCs, per se, to the site of the injury or defect. That is, the wound, injury, or defect is not treated by applying the MPCs within the injury or defect with the idea that those cells will differentiate into the cell types needed to fill the gap or eliminate the defect. Rather, they are placed proximate, but outside, the actual defect, and in fluid communication with the defect, such that the biologically active agents and factors can migrate to the defect and promote the body's natural healing and/or regeneration processes. By promoting those processes, injuries or defects are healed more promptly, and injuries that might not otherwise be able to heal at all are effectively treated.
- the scaffold component of these grafts generally has a structure including a matrix material that is porous in which the MPCs are seeded.
- the matrix material is made of nonaligned fibers that form many discrete interstices or voids, which, due to the complex topology of the matrix serve to retain the MPCs.
- the MPCs can be implanted within the matrix by passive infusion, by mechanical injection or introduction into the interior of the matrix, or by cultivating the cells to grow within the matrix, or some combination thereof. Additionally, the cells can be fixed more stringently within the matrix by chemical modification of the matrix to increase the coefficient of friction between the matrix and the cells, or by increasing the chemical attraction or bonding (e.g., covalent or ionic) between the matrix and the MPCs.
- the matrix is highly adaptable, and can be modified to whatever shape and dimension is best suited to put the MPCs proximate and in fluid contact with the injury or defect.
- the MPC-seeded matrix can be in the form of a patch, wrap, conduit, or the like.
- patches and wraps can be further modified and configured to fit the contours and dimension of the wound as in any bandage or wound treatment.
- the matrix can be further modified in accordance with known principles to be used internally or externally.
- the shape and dimension of the matrix can be further modified to complement other structural features of the graft.
- the matrix can be made to serve as an outer sheath of a conduit, wrapping around, enveloping, or overlaying structural features that might serve as scaffolding for tissue growth.
- a graft comprising a porous matrix of non-aligned fibers forming interstices; the interstices consisting of a cellular component and a non-cellular component, and wherein the majority of the cellular component is MPCs; a conduit internal to the matrix; and wherein the interior of the conduit is in fluid communication with the matrix.
- MPCs mobile cellular component
- conduit internal to the matrix
- the interior of the conduit is in fluid communication with the matrix.
- the matrix, aligned fibers, conduit, or other elements of the grafts disclosed herein can be formed of any known and adaptable biocompatible structural material that lends itself to fabrication according to the demands of the end use.
- the material is a synthetic structural material such as a biocompatible polymer.
- Biodegradable polymers are also desirable. Those materials can be selected such that they are dissolved or resorbed by the body without the need for surgical removal procedures.
- Biocompatible, biodegradable materials useful in the grafts disclosed herein include polyglycolic acid (PGA), type 1 collagen, Poly-DL-lactidecaprolactone (PCL), laminin, gelatin, and the like.
- the graft may further comprise a liquid impermeable exterior liner overlaying the matrix.
- the impermeable exterior liner isolates the fluid and cells of the interior of the conduit with the external media.
- Impermeable generally refers to fluid impermeable, though it also contemplates a less than perfectly fluid impermeable membrane.
- the exterior liner can be used to seal the interior of the graft. This will be advantageous in vascular grafts.
- the exterior liner can also be configured to include additional structural elements such as flanges, sleeves, connectors, and other sealing means. Those additional structural elements can be configured to create a connection or, alternatively, a seal between the biological conduits of the vasculature and the conduit of the graft, and to isolate the contents of the two from external biological media.
- Connecting elements used to effect a seal or connection between the graft and vasculature (or other conduit) can be fabricated from the same material as the impermeable exterior liner, or may be fabricated of another material, and may be part of yet another layer or structural element altogether.
- the nerve tube comprises several chambers or sheaths (e.g., two or more).
- the tube may comprise two sheaths, which may in turn be surrounded or encased within a conduit wall, which may optionally provide a fluid-impermeable exterior wall thus isolating the nerve tube and its contents from the exterior environment.
- the inner sheath comprises aligned fibers generally parallel to a longitudinal axis of symmetry through the tube.
- the fibers may be nanofibers.
- the sheath of aligned fibers is surrounded by another sheath containing random or non-aligned and/or non-linear fibers.
- the inner and outer sheaths may be in fluid communication.
- the exterior wall may extend in length beyond the inner and/or outer sheaths.
- the tube may be placed around a proximal and/or distal nerve stump, optionally enclosing and isolating a space between the stumps.
- the nanofibers are oriented within an inner sheath at the core of a generally tubular structure having an inner sheath and an outer sheath.
- the tubular structure may be circular or elliptical, or any shape necessary to align the nerve tube with the injured or damaged nerves needing repair or regeneration.
- the inner sheath constitutes aligned fibers paralleling an axis of symmetry through the center of a tube.
- the aligned fibers may be linear, parallel fibers less than about 100 ⁇ m, or less than about 10 ⁇ m, and running the length of the tube.
- the nanofibers create sub-micron sized scaffolding that provides a structural framework supporting the growth of nerve regeneration elements, e.g., axons, along the length of the nerve tube or graft.
- the aligned fibers provide three dimensional support and directional orientation for the growing axon.
- the aligned fibers may be made of a biocompatible, biodegradable material that is resorbed or dissolved within the body avoiding the need for surgical removal.
- the nerve tube or graft device further includes a three-dimensional matrix in an outer sheath of the graft.
- the outer sheath of the nerve tube or graft device surrounds the aligned nanofibers, and may permit fluid exchange between the two.
- the outer sheath may share an axis of symmetry with the nerve tube, and may be generally concentric with the bundle of aligned fibers of the inner sheath.
- aligned fibers is meant that the fibers are generally aligned along a longitudinal axis of the inner sheath or tube. The fibers are not necessarily strictly parallel, and some deviation in direction and linearity is contemplated.
- the material of the outer sheath provides a support matrix for multipotent cells, particularly MPCs, within the outer sheath.
- the multipotent cells can be MPCs alone or in combination with MSCs and/or other regenerative cells.
- the nerve tube comprises a tube comprising two zones or sheaths.
- the zones or sheaths may be coaxial.
- the inner sheath may comprise aligned fibers generally parallel to an axis of symmetry along the length of the nerve tube. The ends of the aligned fibers are exposed to the exterior of the tube permitting axonal growth along the fibers through the tube.
- an outer sheath Surrounding the inner sheath, and optionally in fluid contact therewith, is an outer sheath comprising non-aligned and/or non-linear fibers.
- the fibers of the two sheaths may be of the same or different material, and may or may not be of the same size, shape, diameter, and dimension.
- Either or both of the sheaths of the nerve tube may further comprise biologically active agents in addition to those secreted by MSCs and/or MPCs, e.g., hormones, steroids, anti-inflammatories, analgesics, immunosuppressives, anticoagulants, muscle relaxants and antispasmodics, antibiotics and/or antimicrobials, growth factors, colony stimulating factors, nutrients such as vitamins, peptides, small drug molecules, gene therapy agents, e.g., plasmids, retrovirals, and combinations thereof; and other pharmacologically acceptable excipients, diluents, buffers, preservatives, and the like.
- Either or both of the sheaths may further comprise growth media and/or nutrients, antibiotics, preservatives, buffers, and the like to maintain viability of the MSCs and/or MPCs. Other multipotent cells may also be included.
- Also disclosed is a method of forming a nerve graft in a patient suffering an injury to a nerve comprising: seeding a nerve guide or tube with MPCs; and implanting the nerve guide between a proximal nerve stump and a distal nerve stump.
- the implant may be removed following regeneration of the nerve; or the implant may be made of a resorbable material eliminating the need of a surgical removal process.
- the nerve guide is constructed and/or implanted such that it resides proximate to the nerve stumps, and bridging the gap between the nerve stumps. Thus, each end of the nerve guide or tube connects the nerve.
- the nerve guide or tube is maintained in position relative to the nerve stumps for a period sufficient to guide axonal growth and restoration between the respective nerve stumps.
- the term "nerve stump” refers to the one or more residual portions of a nerve following injury or damage to the nerve resulting in a loss of function, e.g., as by severed axon. Nerve injury may be in the form of traumatic injury, surgery, the result of disease, or other cause.
- the nerve guide is a peripheral nerve guide as disclosed elsewhere herein.
- the guide can be used to bridge a critical sized defect in a peripheral nerve following traumatic injury.
- the device has at least two components (see, e.g ., Figure 1 ).
- the scaffold can be fabricated using polycapralactone, or other suitable biocompatible and/or biodegradable polymer such as polylactic acid, collagen, laminin, gelatin, and the like.
- the polymer is formed into nano-meter scale fibers (e.g., about 50-500 nm diameter; or about 100-300 nm diameter; or about 200 nm diameter).
- the scaffold is formed by extrusion of polymer using an electrospinning process.
- electrospinning process it will be appreciated by those skilled in the art that other methods of fabricating such fibers are available, and thus are contemplated herein.
- the scaffold contains a core of aligned nanofibers, which guide and support growing axons to bridge the defect in the nerve.
- the aligned fibers may be surrounded by a sheath of non-aligned, randomly oriented fibers.
- MPCs may be seeded in the sheath of non-aligned fibers.
- MPCs can be cultured to secrete biochemical factors. Those factors enhance the growth rate of axons, and recruit native neuronal support cells into the device.
- MPCs can be used as the only active growth promoting agent within the scaffold, or in combination with other agents, including other multipotent cells such as stem cells, including MSCs.
- the instant scaffold or grafting device is not limited to nerve grafts, but may be used to repair various injuries including bone, blood vessel, muscle, ligament, and tendon.
- the grafts disclosed herein assist nerve regeneration. They provide a protective environment for axonal sprouting and improve the chances that the damaged axons will reach the distal nerve stump.
- Biocompatible, biodegradable materials useful in the grafts disclosed herein include polyglycolic acid (PGA), type 1 collagen, Poly-DL-lactidecaprolactone (PCL).
- the instant grafts have the particular advantage of providing individual axon guidance with nanofiber filaments, and facilitate access and activity of axonal support cells such as Schwann cells or MSCs. Both of these features constitute improvements over currently available devices, and enhance the speed of axonal growth. The speed and accuracy of axonal growth determines the success of reinnervation.
- the instant devices provide improved regeneration of peripheral nerves compared to the currently available alternatives. Furthermore, by increasing the speed of axonal growth, the instant nerve guides and methods expand the versatility and range of nerve regeneration; and enable regeneration of nerves suffering defects of substantially greater size than has been possible under prior therapies.
- the first device comprises a solid core of aligned nanofibers.
- the nanofibers support growth of regenerating axons.
- aligned fibers have been shown to accelerate the rate of axonal growth, such use of fibers has been limited.
- a technique used to fabricate a sheet of aligned fibers has been reported, and previous investigators have cut strips of these aligned fibers and pulled them through the lumen of a conventional nerve guide. Those strips or sheets of aligned fibers afford only a two dimensional framework, and diminishes migration of Schwann cells into the lumen. That has the effect of diminishing growth of the axon through the lumen, and less effective regeneration of the nerve.
- all of the regenerating axons have access to aligned nanofibers. This can be effected by surgically implanting the device so that it is contact with the two nerve stumps.
- the device is formed by electrospinning the non-aligned sheath of nanofibers that surround the aligned core ( Figure 2 ).
- electrospinning is performed by passing a polymer solution through a blunt-tipped needle that contains a high electric charge (e.g., greater than about 15kV). As the solution passes through the needle, part of the charge is transferred to the polymers, which are extruded out of the needle towards a grounding plate.
- the grounding plate is replaced with two axels that are aligned end-on. In between the axels, there is a strip of aligned nanofibers, and the entire assembly is rotated as a mandrel.
- the resulting nanofibers are attracted to ground on one of the two axels, and the target axel can switch stochastically.
- the fiber is stretched over the strip of aligned nanofibers.
- the aligned nanofibers become completely encased by the sheath of non-aligned fibers.
- a graft for use in a method of treating a patient suffering an injury to tissue and requiring restoration and/or regeneration of the tissue comprising applying to the injury a graft comprising a porous matrix of non-aligned fibers forming interstices, the interstices consisting of a cellular component and a non-cellular component, and wherein the majority of the cellular component is mesenchymal progenitor cells.
- interstices refers to voids among the fibers.
- the voids can be seeded with, or infused with, MPCs and other cell types, cell culturing components, and non-cellular components such as other active ingredients. Use of the term majority means greater than 50%.
- MPCs can be seeded within the graft such that the majority of MPCs are not in direct contact with the tissue requiring repair or regeneration.
- the treatment method contemplates that the MPCs are placed in proximity to the injury or damage site, but are not positioned or administered in such a way that they are used to pack a gap or defect in the tissue. That is, the MPC seeded matrix will not form a structural element restoring or regenerating the damaged or missing structural or function elements of the wounded tissue, but will be part of a separate matrix that is not ultimately incorporated into the gap or defect.
- the MPC-seeded matrixes described herein will be a biocompatible, biodegradable material that is dissolved or resorbed by the body, but preferably not until the gap or defect in the tissue is healed or sufficiently diminished that the body's natural healing capacity can take over and complete the healing and/or regeneration process.
- the methods and materials disclosed herein can be used to: prevent fibrosis; augment muscle regeneration; improve fracture healing.
- Devices can also be seeded with MPCs for tissue engineering, e.g., peripheral nerve; bone; blood vessel; tendons and ligaments.
- MSCs Mesenchymal Progenitor cells
- tissue specimens were obtained from patients who had sustained traumatic extremity injury during Operation Iraqi Freedom and Operation Enduring Freedom. These patients presented to Walter Reed Army Medical Center approximately three to seven days after the injury and underwent serial debridement and irrigation procedures until the wounds were determined to be clinically acceptable for definitive orthopaedic treatment.
- the amount and nature of debrided tissue was surgeon dependent and was based on trauma surgery principles of circumferential removal of all grossly contaminated, apparently necrotic, and nonviable tissue along with a thin margin of healthy-appearing tissue. This procedure was repeated at each surgical encounter until only healthy tissue remained, and cells typically were harvested from muscle tissue obtained during the second or third serial debridement.
- the protocol for extracting muscle-derived multiprogenitor cells was based on a modification of previous work in isolating mesenchymal stem cells that was performed in our laboratory. Caterson EJ, Human marrow-derived mesenchymal progenitor cells: isolation, culture expansion, and analysis of differentiation. Mol Biotechnol. 2002;20:245-56 . Fat, fascia, other connective tissue, and necrotic tissue were dissected away from the healthy margin of the debrided muscle sample. Approximately 0.5 cc of the remaining muscle tissue was processed for cell extraction.
- the tissue was washed three times in Hanks' Balanced Salt Solution (Gibco, Carlsbad, California) and then was extensively minced in a 10-cm culture dish containing Dulbecco's Modified Eagle Medium (Gibco) and 3X penicillin/streptomycin/Fungizone (Gibco) until it could pass through the tip of a 25-mL serological pipette (Falcon; BD Biosciences, San Jose, California).
- Hanks' Balanced Salt Solution Gibco, Carlsbad, California
- the minced tissue was transferred to a 50-mL conical vial with digestion medium containing Dulbecco's Modified Eagle Medium, 3X penicillin/streptomycin/Fungizone, and 0.5 mg/mL collagenase type 2 (Worthington Biochemical, Lakewood, New Jersey).
- tissue slurry was agitated gently at 37°C for two hours, and the resulting digest was filtered through a 40- ⁇ m cell strainer (Falcon), pelleted by means of centrifugation, resuspended in growth medium (Dulbecco's Modified Eagle Medium with 10% fetal bovine serum; Gibco) and 5X penicillin/streptomycin/Fungizone, and then plated onto tissue culture polystyrene (150-cm 2 flask; Falcon).
- growth medium Dulbecco's Modified Eagle Medium with 10% fetal bovine serum; Gibco
- 5X penicillin/streptomycin/Fungizone tissue culture polystyrene (150-cm 2 flask; Falcon).
- the cells were incubated at 37°C in a 5% CO 2 -humidified cell incubator for two hours and then were extensively washed with Hanks' Balanced Salt Solution before fresh growth medium was added with 3X penicillin/streptomycin/Fungizone. Once multiprogenitor cell colony forming units were observed, the concentration of penicillin/streptomycin/Fungizone was lowered to 1X. Cell confluence was obtained after approximately two weeks. The cell cultures were routinely passaged at 80% to 90% confluence and split 1:4.
- MPCs express higher levels of COL15A1, a gene associated with muscle tissue development, and GDF10, shown to be a negative regulator of osteogenesis, whereas the bone-marrow derived MSCs express higher levels of genes associated with bone physiology and maintenance: VEGFA, VCAM1 and IGF2. These differences may also reflect the fact that traumatized muscle-derived MPCs are harvested from an active wound bed, where they likely participate in the process of muscle tissue repair.
- COL15A1 and GDF10 are substantially, albeit non-significantly, down-regulated, while VEGFA, VCAM1 and IGF2 are similarly up-regulated, suggesting that the MPCs can assume the role of osteoprogenitors under the appropriate biological environment, in a manner similar to other populations of MSCs.
- FGF10 is a gene associated with development and the initiation of wound healing
- GDF6 codes for a cytokine that works in concert with bone Morphogenic proteins
- VCAM1 provides a molecular adhesion to vascular structures. The VCAM1 result corroborates the finding from the previous experiment.
- These genes may play a role in the ability of MSCs to reside in the bone marrow and detect damage to the bone.
- Mouse anti-CD29 monoclonal IgG (clone Ha2/5), mouse anti-CD44 monoclonal IgG (clone IM7), mouse anti-CD105 monoclonal IgG (clone 35), and mouse anti-CD146 monoclonal IgG (clone P1H12) antibodies and phytoerythrin-conjugated mouse anti-CD45 monoclonal IgG (clone TU116), mouse anti-CD73 monoclonal IgG (clone AD2), mouse anti-CD90 monoclonal IgG (clone 5E10), and mouse anti-CD105 monoclonal IgG (clone 35) antibodies were obtained from BD Biosciences (San Jose, California). All antibodies were reactive against human antigens. Donkey anti-mouse IgG conjugated with fluorescein isothiocyanate were obtained from Jackson ImmunoResearch (West Grove, Pennsylvania). Testing with negative and positive controls confirmed the specificity of these antibodies.
- Cells used for staining were cultured in growth medium on glass coverslips for fourteen days during the second or third passage. They were washed once with Hanks' Balanced Salt Solution and then were fixed in 3% phosphate-buffered paraformaldehyde for twenty minutes. Fixed cells were first blocked in 2% bovine serum albumin (Sigma-Aldrich, St.
- the coverslips were then mounted to slides with VECTASHIELD (Vector Laboratories, Burlingame, California) and viewed with a Zeiss 510 Metaconfocal laser scanning microscope (Carl Zeiss Microimaging, Thornwood, New York).
- the cells were incubated in the dark at 4°C for forty minutes, washed once in fluorescence-activated cell-sorting buffer, and resuspended in 100 ⁇ L of fresh fluorescence-activated cell sorting buffer.
- the fluorescent intensity profiles of the cells were analyzed by means of fluorescence-activated cell-sorting with use of a FACSCalibur flow cytometer (BD Biosciences).
- the MPCs were positive for CD44, CD49e, CD73, CD90 and CD105 and negative for CD14, CD31, CD34 and CD45.
- Example 3 MPC in vitro expression of trophic factors
- the progenitor cells derived from traumatized muscle were exposed to defined glial-induction media.
- Conditions to optimize the neurotrophic potential of MSCs and muscle-derived progenitor cells were determined using ELISAs to measure the concentration of secreted neurotrophic factors (i.e., BDNF: Brain Derived Neurotrophic Factor, NGF: Nerve Growth Factor, GDNF: Glial Derived Neurotrophic Factor, etc,).
- BDNF Brain Derived Neurotrophic Factor
- NGF Nerve Growth Factor
- GDNF Glial Derived Neurotrophic Factor
- the cells were capable of producing substantial amounts of neurotrophic factors, even without neuroglial induction. After 7 days in defined conditions for glial differentiation, the progenitor cells began to produce neurotrophic factors. In particular, the progenitor cells could produce substantial amounts of BDNF when cultured under optimal induction conditions. Evidence also suggests that they expressed Glial Fibullary Acid Protein, a glial cell specific marker. The amount of CNTF produced by the progenitor cells was unaffected by the neuroglial induction media. MPCs may also express nestin following neurotrophic induction, and the percent of nestin positive cells appears to increase following neurotrophic induction.
- RA retinoic acid
- BME ⁇ -mercaptoethanol
- the final system of neurotrophic induction includes pre-treatment with RA, followed by 7 days in the optimized neurotrophic induction media.
- This system increased the production of BDNF and two other neurotrophic factors important to peripheral nerve regeneration: CNTF and NT-3 ( Figure 7 ).
- Table 1 Formulations for the Neuroglial-Induction Media Media Formulation GM DMEM with 10% FBS NM0 Neurobasal Medium with 2% B27 Supplement NM1 Neurobasal Medium with 2% B27 Supplement, 5 ⁇ M cAMP, 5 ⁇ M IBMX, 2.5 ⁇ g/mL Insulin and 25 ng/mL NGF NM2 Neurobasal Medium with 2% B27 Supplement, 10 ng/mL bFGF, 20 ng/mL EGF and 10 ng/mL of LIF NM3 ⁇ MEM with 10% FBS, 5% Horse Serum, 50 ⁇ M Hydrocortisone and 0.1 ⁇ m Dexamethasone NM4 DMEM/HAMS F12 with 2% B27 Supplement, 2% FBS, 20 ⁇ M Retinoic Acid and 10 ng/mL bFGF
- FGF2 and TGFB3 are somewhat general cytokines that promote growth and scarless wound healing, respectively.
- HGF, LIF and IL10 promote immunosuppression.
- VEGFA promotes vascular regeneration, and BMP2, BMP4 and BMP6 have been shown to promote bone regeneration.
- the MPCs produced a greater amount of VEGFA at days 1, 2 and 4 than MSCs than MSCs, as observed with western blots ( Figure 9 ).
- MMPs play a role in promoting endothelial cell migration and infiltration. Similar to MSCs, MPCs express MMP-2 and MMP-9. Comparison of the two cell types indicates they express similar levels. However, MPCs begin expressing higher levels of MMP-2 and MMP-9 earlier, i.e. by Day 1 ( Figure 10 ).
- This experiment was performed in two ways. First, in a conditioned media experiment, we cultured the progenitor cells for three days in either growth or neurotrophic induction medium. Then, we transferred the media to the DRG cultures for an additional three days. For both media types, medium that was conditioned by the MPCs resulted in a higher density of neurite extensions compared to the corresponding no cell controls. Figure 11 . We further quantified this finding by counting the number of extended neurites under each condition and found that the factors secreted by the progenitor cells resulted in a significant increase in the number of neurites that extended beyond the minimum neurite length.
- MPCs were allowed to secrete their trophic factors into conditioned medium for three days. This medium was then added to fresh media and transferred to endothelial cell culture. Substantial increase in the proliferation of the endothelial cells was observed, as a result of the trophic factors secreted by the MPCs ( Figure 12 ).
- a mixed lymphocyte reaction was performed to evaluate the immunosuppressive properties of the MPCs.
- the proliferation to T cells was measured following stimulation with an antigen.
- the factors secreted by MPCs significantly decreased the T-cell proliferation in a dose-dependent manner ( Figure 13 ).
- a novel, composite electropsun nanofiber scaffold was fabricated and seeded with the MPC cell population to produce a peripheral nerve graft.
- the composite scaffold was fabricated by electrospinning poly( ⁇ )caprolactone into nanometer-scale fibers ( Figures 1 and 2 ).
- the interior core of the scaffold is filled with aligned nanofibers ( Figure 14a ), which are designed to guide axon growth along the interior of the scaffold structure; (2) the aligned fibers of the scaffold are surrounded by a core of non-aligned fibers that support the seeded progenitor cells; and (3) these cells will secrete their neurotrophic factors into the interior core of the scaffold to augment the nerve regeneration process.
- Assays were performed to determine the biological performance of the MPCs in the 3-D environment of the peripheral nerve graft. Viability of the cells in the graft was assessed using a calcein/ethidium bromide fluorescence assay. Within 24 hours, 98% of the seeded cells remained viable and continued to proliferate on the graft at approximately the same rate ( Figure 14b ). After one week in culture, the nerve graft was fixed and prepared for immunohistochemical evaluation of the neurotrophic factor distribution in the construct. The seeded cells produced BDNF while in the graft in both growth media and neutrophic induction media, with the MPCs cultured in the neurotrophic induction medium producing more BDNF on a per cell basis ( Figure 14c ).
- Figure 14d shows cross sections through the graft, with the dotted line indicating the interface between the non-aligned exterior of the graft and the aligned interior core, and with the aligned fibers running in and out of the figure.
- BDNF was localized near the MPCs in the non-aligned region of the graft.
- the secreted factors also enriched the aligned fibers.
- the MPCs demonstrated the ability to remain viable and functional after seeding onto the composite nanofibrous scaffold of the peripheral nerve graft.
- the neurotrophic factors they produced will enhance the neuroconductivity of the aligned nanofibers.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Organic Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Dermatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Cell Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Rheumatology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Cardiology (AREA)
- Heart & Thoracic Surgery (AREA)
- Virology (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Vascular Medicine (AREA)
- Dispersion Chemistry (AREA)
- Botany (AREA)
Claims (13)
- Greffe pour promouvoir une guérison d'une lésion ou d'un défaut comprenant un échafaudage et un composant thérapeutique, l'échafaudage comprenant une matrice poreuse de fibres non alignées formant des interstices, dans laquelle le composant thérapeutique comprend un composant cellulaire comprenant des cellules progénitrices mésenchymateuses et, facultativement, un composant non cellulaire, dans laquelle les cellules progénitrices mésenchymateuses sont retenues dans les interstices formés par les fibres non alignées de la matrice poreuse de la greffe d'une manière qui place les cellules progénitrices mésenchymateuses à proximité de et en contact de fluide avec, mais à l'extérieur de, la lésion ou le défaut et évite un contact des cellules progénitrices mésenchymateuses avec le site de la lésion ou du défaut, dans laquelle la greffe a la forme d'un conduit comprenant une gaine interne de nanofibres alignées qui est entouré par les fibres non alignées formant une gaine externe.
- Greffe selon la revendication 1, dans laquelle la matrice forme une gaine externe autour d'une zone intérieure et les cellules progénitrices mésenchymateuses sont ensemencées dans la matrice d'une manière qui permet à des facteurs excrétés par les cellules progénitrices mésenchymateuses de migrer dans la zone intérieure.
- Greffe selon la revendication 1, dans laquelle les nanofibres ont un diamètre d'environ 0,05 à 0,5 µm.
- Greffe selon la revendication 1 ou la revendication 2, dans laquelle les fibres sont des fibres synthétiques.
- Greffe selon la revendication 4, dans laquelle les fibres sont des fibres polymères biocompatibles et biodégradables.
- Greffe selon la revendication 1 ou la revendication 2, dans laquelle le composant cellulaire comprend en outre d'autres cellules souches.
- Greffe selon la revendication 1 ou la revendication 2, dans laquelle le composant non cellulaire comprend un milieu de culture cellulaire.
- Greffe selon la revendication 1, dans laquelle le composant non cellulaire comprend des agents actifs sur le plan pharmaceutique choisis dans le groupe constitué de facteurs de croissance, anti-inflammatoires, antibiotiques et antiviraux.
- Greffe selon la revendication 1, dans laquelle le composant non cellulaire comprend des ingrédients actifs sur le plan thérapeutique.
- Greffe selon la revendication 1 ou la revendication 2, comprenant en outre un revêtement extérieur imperméable aux liquides recouvrant la matrice.
- Greffe selon la revendication 1 pour une utilisation dans le traitement d'un patient souffrant d'une lésion à un tissu et exigeant une restauration et/ou une régénération du tissu.
- Greffe pour l'utilisation selon la revendication 11, dans laquelle le tissu exigeant une réparation ou une régénération est choisi dans le groupe constitué de : tissu nerveux, osseux et vasculaire.
- Greffe selon la revendication 2 pour une utilisation dans le traitement d'un patient souffrant d'une lésion nerveuse produisant un espace dans le nerf.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11781408P | 2008-11-25 | 2008-11-25 | |
PCT/US2009/004482 WO2010062297A1 (fr) | 2008-11-25 | 2009-08-05 | Greffes de tissu régénératives et leurs procédés de fabrication |
Publications (3)
Publication Number | Publication Date |
---|---|
EP2358301A1 EP2358301A1 (fr) | 2011-08-24 |
EP2358301A4 EP2358301A4 (fr) | 2013-12-11 |
EP2358301B1 true EP2358301B1 (fr) | 2019-07-17 |
Family
ID=42225950
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP09829422.6A Not-in-force EP2358301B1 (fr) | 2008-11-25 | 2009-08-05 | Greffes de tissu régénératives et leurs procédés de fabrication |
Country Status (5)
Country | Link |
---|---|
US (2) | US8652458B2 (fr) |
EP (1) | EP2358301B1 (fr) |
AU (1) | AU2009320446B2 (fr) |
CA (2) | CA2967583C (fr) |
WO (1) | WO2010062297A1 (fr) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11311366B2 (en) | 2010-06-17 | 2022-04-26 | Washington University | Biomedical patches with aligned fibers |
US11596717B2 (en) | 2012-09-21 | 2023-03-07 | Washington University | Three dimensional electrospun biomedical patch for facilitating tissue repair |
Families Citing this family (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9192695B2 (en) | 2008-11-20 | 2015-11-24 | Allosource | Allografts combined with tissue derived stem cells for bone healing |
CA2967583C (fr) | 2008-11-25 | 2020-09-01 | The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. | Methodes d'obtention de cellules progenitrices mesenchymateuses |
US20140286911A1 (en) | 2013-03-15 | 2014-09-25 | Allosource | Cell repopulated collagen matrix for soft tissue repair and regeneration |
US9162011B2 (en) | 2011-12-19 | 2015-10-20 | Allosource | Flowable matrix compositions and methods |
CA2867167A1 (fr) * | 2012-02-13 | 2013-08-22 | Board Of Regents, The University Of Texas System | Systeme d'echafaudage pour reparation de tissu |
WO2013151725A1 (fr) | 2012-04-05 | 2013-10-10 | The Regents Of The University Of California | Cellules de sérums de régénération et cellules souches mésenchymateuses |
RU2545993C2 (ru) * | 2013-09-05 | 2015-04-10 | Александр Сергеевич Тепляшин | Способ восстановления костных дефектов трубчатых костей критической величины |
CL2013003066A1 (es) | 2013-10-22 | 2014-07-25 | Univ Chile | Composicion para tratamiento de heridas porque comprende matriz soporte y células troncalesmesenquiámicas de gelatina de wharton; metodo para tratar heridas que comprende aplicar dicha composicion |
US9849219B2 (en) * | 2014-03-14 | 2017-12-26 | Dermagenesis, Llc | Methods for harvesting human fibroblasts, and autografting methods |
EP3229739A4 (fr) * | 2014-12-09 | 2018-10-31 | Rutgers, the State University of New Jersey | Échafaudage tridimensionnel pour régénération osseuse |
CN104623738B (zh) * | 2015-01-12 | 2016-08-24 | 南通大学 | 带有悬丝纤维支架的组织工程神经移植物及其制备方法 |
WO2016192733A1 (fr) * | 2015-05-29 | 2016-12-08 | Aarhus Universitet | Conduit de régénération de matériau biologique |
US10632228B2 (en) | 2016-05-12 | 2020-04-28 | Acera Surgical, Inc. | Tissue substitute materials and methods for tissue repair |
PL234234B1 (pl) * | 2016-12-22 | 2020-01-31 | Inst Biopolimerow I Wlokien Chemicznych | Proteza z rdzeniem chitozanowym do regeneracji nerwów i sposób jej wytwarzania |
IT201700064613A1 (it) * | 2017-06-12 | 2018-12-12 | Univ Bologna Alma Mater Studiorum | Scaffold multiscala elettrofilato per la rigenerazione e/o sostituzione del tessuto tendineo/legamentoso e metodo di produzione |
CN111097069A (zh) * | 2019-07-04 | 2020-05-05 | 南开大学 | 一种利用拓扑结构调控免疫微环境并引导再生的仿生可降解人工神经导管及其制备方法 |
GB202000714D0 (en) | 2020-01-17 | 2020-03-04 | Ucl Business | Biomaterials |
US11957815B2 (en) | 2020-07-17 | 2024-04-16 | Datt Life Sciences Private Limited | Ready to use biodegradable and biocompatible cell-based nerve conduit for nerve injury and a method of preparation thereof |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US111390A (en) * | 1871-01-31 | Improvement in heaters for paper-ruling machines | ||
AU7611500A (en) * | 1999-09-24 | 2001-04-24 | Abt Holding Company | Pluripotent embryonic-like stem cells, compositions, methods and uses thereof |
GB0030583D0 (en) | 2000-12-15 | 2001-01-31 | Univ Nottingham | Nerve regeneration |
WO2006113642A1 (fr) * | 2005-04-18 | 2006-10-26 | Duke University | Structure en fibres tridimensionnelles pour des techniques associees a des tissus specifiques |
WO2006138552A2 (fr) | 2005-06-15 | 2006-12-28 | Government Of The Usa, As Represented By The Secretary, Dept. Of Health & Human Services | Cartilage a tissu modifie, procede de fabrication correspondant et applications chirurgicales therapeutiques et cosmetiques qui l'utilisent |
DE102005054941A1 (de) * | 2005-11-17 | 2007-05-31 | Gelita Ag | Nervenleitschiene |
EP2599858A3 (fr) * | 2006-01-27 | 2013-09-18 | The Regents of The University of California | Squelettes biomimétiques |
WO2007146106A2 (fr) * | 2006-06-05 | 2007-12-21 | Cryo- Cell International, Inc. | Obtention, isolation et cryoconservation de cellules placentaires maternelles |
CA2967583C (fr) | 2008-11-25 | 2020-09-01 | The Henry M. Jackson Foundation For The Advancement Of Military Medicine, Inc. | Methodes d'obtention de cellules progenitrices mesenchymateuses |
-
2009
- 2009-08-05 CA CA2967583A patent/CA2967583C/fr not_active Expired - Fee Related
- 2009-08-05 AU AU2009320446A patent/AU2009320446B2/en not_active Ceased
- 2009-08-05 EP EP09829422.6A patent/EP2358301B1/fr not_active Not-in-force
- 2009-08-05 WO PCT/US2009/004482 patent/WO2010062297A1/fr active Application Filing
- 2009-08-05 US US13/129,636 patent/US8652458B2/en not_active Expired - Fee Related
- 2009-08-05 CA CA2744559A patent/CA2744559C/fr not_active Expired - Fee Related
-
2014
- 2014-02-12 US US14/178,932 patent/US10507187B2/en not_active Expired - Fee Related
Non-Patent Citations (1)
Title |
---|
MAHAY DALJEET ET AL: "Schwann cell mediated trophic effects by differentiated mesenchymal stem cells.", EXPERIMENTAL CELL RESEARCH 15 AUG 2008, vol. 314, no. 14, 15 August 2008 (2008-08-15), pages 2692 - 2701, XP023783389, ISSN: 1090-2422 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11311366B2 (en) | 2010-06-17 | 2022-04-26 | Washington University | Biomedical patches with aligned fibers |
US11471260B2 (en) | 2010-06-17 | 2022-10-18 | Washington University | Biomedical patches with aligned fibers |
US11596717B2 (en) | 2012-09-21 | 2023-03-07 | Washington University | Three dimensional electrospun biomedical patch for facilitating tissue repair |
Also Published As
Publication number | Publication date |
---|---|
CA2967583C (fr) | 2020-09-01 |
EP2358301A4 (fr) | 2013-12-11 |
WO2010062297A1 (fr) | 2010-06-03 |
US10507187B2 (en) | 2019-12-17 |
CA2744559C (fr) | 2017-07-04 |
AU2009320446A1 (en) | 2011-07-07 |
AU2009320446B2 (en) | 2015-08-20 |
US8652458B2 (en) | 2014-02-18 |
EP2358301A1 (fr) | 2011-08-24 |
CA2744559A1 (fr) | 2010-06-03 |
US20110318414A1 (en) | 2011-12-29 |
CA2967583A1 (fr) | 2010-06-03 |
US20140227339A1 (en) | 2014-08-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP2358301B1 (fr) | Greffes de tissu régénératives et leurs procédés de fabrication | |
US11179499B2 (en) | Bone-like prosthetic implants | |
Kuhbier et al. | Isolation, characterization, differentiation, and application of adipose-derived stem cells | |
Kang et al. | Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton's jelly for treating bone defects | |
US10675307B2 (en) | Compositions comprising perivascular stem cells and nell-1 protein | |
Kim et al. | Intra-articular delivery of synovium-resident mesenchymal stem cells via BMP-7-loaded fibrous PLGA scaffolds for cartilage repair | |
WO2014114043A1 (fr) | Greffe de nerf par ingénierie tissulaire avec modification de matrice cellulaire pour réparation de lésion de nerf périphérique et son procédé de préparation | |
WO2007016621A2 (fr) | Utilisation de cellules stromales derivees de tissu adipeux dans une fusion spinale | |
Espitalier et al. | A comparison between bone reconstruction following the use of mesenchymal stem cells and total bone marrow in association with calcium phosphate scaffold in irradiated bone | |
de Assis et al. | Stem cells and tissue engineering-based therapeutic interventions: promising strategies to improve peripheral nerve regeneration | |
Trombi et al. | Human autologous plasma‐derived clot as a biological scaffold for mesenchymal stem cells in treatment of orthopedic healing | |
KR20200000823A (ko) | 인간 유도 만능 줄기세포로부터 연골세포의 펠렛을 제조하는 방법 및 이의 용도 | |
Mao et al. | Demineralized cortical bone matrix augmented with peripheral blood-derived mesenchymal stem cells for rabbit medial meniscal reconstruction | |
da Rocha et al. | A new option for bone regeneration: a rapid methodology for cellularization of allograft with human bone marrow stromal cells with in vivo bone-forming potential | |
KR101649375B1 (ko) | 세포이식술을 위한 혼합세포복합체인 세포스페로이드의 제조방법 및 이의 이용방법 | |
AU2019203773A1 (en) | Regenerative tissue grafts and methods of making same | |
Catalano et al. | Adipose-derived adult stem cells: available technologies for potential clinical regenerative applications in dentistry | |
Fan et al. | Enhancing synovial mesenchymal stem cell adhesion and selection via an avidin-biotin-CD105 binding system for cartilage tissue engineering | |
WO2016172004A1 (fr) | Système de cicatrisation osseuse, d'accélération de l'angiogenèse et de production de vasculogenèse | |
Xu et al. | Constructing Injectable Bone-Forming Units by Loading a Subtype of Osteoprogenitors on Decellularized Bone Matrix Powders for Bone Regeneration | |
Suh et al. | Effects of co-culture of dental pulp stem cells and periodontal ligament stem cells on assembled dual disc scaffolds | |
Zhu et al. | Harnessing three-dimensional porous chitosan microsphere embedded with adipose-derived stem cells to promote nerve regeneration | |
Hofer | Traumatized muscle-derived multipotent progenitor cells: Pro-angiogenic activity, promotion of nerve growth, and osteogenic differentiation | |
WO2021246893A1 (fr) | Sphéroïde biocomposite pour la restauration des os | |
Yang | Development of Hydrodynamically Engineered Cartilage in Response to Insulin-like Growth Factor-1 and Transforming Growth Factor-beta1: Formation and Role of a Type I Collagen-based Fibrous Capsule |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 20110620 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK SM TR |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: JACKSON, WESLEY, M. Inventor name: TUAN, ROCKY, S. Inventor name: NESTI, LEON, J. |
|
RAP1 | Party data changed (applicant data changed or rights of an application transferred) |
Owner name: THE UNITED STATES GOVERNMENT AS REPRESENTED BY THE Owner name: THE UNITED STATES GOVERNMENT, AS REPRESENTED BY TH Owner name: THE HENRY M. JACKSON FOUNDATION FOR THE ADVANCEMEN |
|
DAX | Request for extension of the european patent (deleted) | ||
A4 | Supplementary search report drawn up and despatched |
Effective date: 20131113 |
|
RIC1 | Information provided on ipc code assigned before grant |
Ipc: A61L 27/18 20060101ALI20131107BHEP Ipc: A61L 27/34 20060101ALI20131107BHEP Ipc: A61F 2/02 20060101AFI20131107BHEP Ipc: A61L 27/38 20060101ALI20131107BHEP Ipc: A61L 27/56 20060101ALI20131107BHEP Ipc: C12N 5/0775 20100101ALI20131107BHEP Ipc: A61L 27/58 20060101ALI20131107BHEP |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20161209 |
|
GRAP | Despatch of communication of intention to grant a patent |
Free format text: ORIGINAL CODE: EPIDOSNIGR1 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: GRANT OF PATENT IS INTENDED |
|
INTG | Intention to grant announced |
Effective date: 20190208 |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: NESTI, LEON, J. Inventor name: TUAN, ROCKY, S. Inventor name: JACKSON, WESLEY, M. |
|
GRAS | Grant fee paid |
Free format text: ORIGINAL CODE: EPIDOSNIGR3 |
|
GRAA | (expected) grant |
Free format text: ORIGINAL CODE: 0009210 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE PATENT HAS BEEN GRANTED |
|
AK | Designated contracting states |
Kind code of ref document: B1 Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK SM TR |
|
REG | Reference to a national code |
Ref country code: GB Ref legal event code: FG4D |
|
REG | Reference to a national code |
Ref country code: CH Ref legal event code: EP |
|
REG | Reference to a national code |
Ref country code: IE Ref legal event code: FG4D |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R096 Ref document number: 602009059174 Country of ref document: DE |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: REF Ref document number: 1155131 Country of ref document: AT Kind code of ref document: T Effective date: 20190815 |
|
REG | Reference to a national code |
Ref country code: NL Ref legal event code: MP Effective date: 20190717 |
|
REG | Reference to a national code |
Ref country code: LT Ref legal event code: MG4D |
|
REG | Reference to a national code |
Ref country code: AT Ref legal event code: MK05 Ref document number: 1155131 Country of ref document: AT Kind code of ref document: T Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: NO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191017 Ref country code: PT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191118 Ref country code: NL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: LT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: AT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: BG Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191017 Ref country code: FI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: HR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: SE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: ES Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: GR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191018 Ref country code: IS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20191117 Ref country code: LV Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: TR Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: DK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: EE Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: PL Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: RO Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: LI Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20190831 Ref country code: MC Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: CH Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20190831 Ref country code: IS Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20200224 Ref country code: SK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: LU Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20190805 Ref country code: CZ Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: SM Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
REG | Reference to a national code |
Ref country code: BE Ref legal event code: MM Effective date: 20190831 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R097 Ref document number: 602009059174 Country of ref document: DE |
|
PLBE | No opposition filed within time limit |
Free format text: ORIGINAL CODE: 0009261 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT |
|
PG2D | Information on lapse in contracting state deleted |
Ref country code: IS |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: IE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20190805 |
|
26N | No opposition filed |
Effective date: 20200603 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: BE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20190831 Ref country code: SI Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PGFP | Annual fee paid to national office [announced via postgrant information from national office to epo] |
Ref country code: FR Payment date: 20200825 Year of fee payment: 12 Ref country code: DE Payment date: 20200827 Year of fee payment: 12 Ref country code: GB Payment date: 20200827 Year of fee payment: 12 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: CY Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MT Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 Ref country code: HU Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT; INVALID AB INITIO Effective date: 20090805 |
|
REG | Reference to a national code |
Ref country code: DE Ref legal event code: R119 Ref document number: 602009059174 Country of ref document: DE |
|
GBPC | Gb: european patent ceased through non-payment of renewal fee |
Effective date: 20210805 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: MK Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT Effective date: 20190717 |
|
PG25 | Lapsed in a contracting state [announced via postgrant information from national office to epo] |
Ref country code: GB Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20210805 Ref country code: FR Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20210831 Ref country code: DE Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES Effective date: 20220301 |